Biocompatible devices, systems, and methods for reducing levels of pro-inflammatory or anti-inflammatory stimulators or mediators in the blood

ABSTRACT

Devices, systems, and methods reduce levels of pro-inflammatory or anti-inflammatory stimulators or mediators in blood by selective adsorption. The devices, systems, and methods are useful in situations where abnormal levels of or unregulated or excessive interaction among pro-inflammatory or anti-inflammatory stimulators or mediators occur, or during events that do induce or have the potential for inducing abnormal production of pro-inflammatory or anti-inflammatory stimulators or mediators. The devices, systems, and methods serve to prevent, control, reduce, or alleviate the severity of the inflammatory response and disease states that are associated with abnormal levels of or unregulated or excessive interaction among pro-inflammatory or anti-inflammatory stimulators or mediators.

RELATED APPLICATIONS

This application is a continuation of U.S. patent application Ser. No.11/255,132, filed Oct. 19, 2005 now abandoned, and entitled “System forTreating Patient with Bacterial Infections,” which is a division of U.S.patent application Ser. No. 10/036,758, filed Dec. 21, 2001 nowabandoned, which is a continuation-in-part of U.S. patent applicationSer. No. 09/832,159, filed Apr. 10, 2001, now abandoned, and entitled“System for Treating Patient with Bacterial Infections,” and which isalso a continuation-in-part of U.S. patent application Ser. No.09/829,252, filed Apr. 10, 2001, now abandoned, and entitled “Method ofTreating Patient with Bacterial Infections,” and which is also acontinuation-in-part of U.S. patent application Ser. No. 09/294,224,filed Apr. 19, 1999, (now U.S. Pat. No. 6,416,487) and entitled “Methodfor Removing Beta-2 Microglobulin from Blood,” which is acontinuation-in-part of U.S. patent application Ser. No. 08/902,727,filed Jul. 30, 1997 (now U.S. Pat. No. 5,904,663), all of which areincorporated herein by reference.

FIELD OF THE INVENTION

This invention relates to devices, systems, and methods for removingtargeted proteins or toxins from the blood, blood products, orphysiologic fluids.

BACKGROUND OF THE INVENTION

In animals, an inflammatory response occurs when tissues are injured bybacteria, trauma, toxins, heat, or other agents, which can becollectively referred to as “Inflammatory Agents.” The nature andcharacter of a given inflammatory response is regulated by the complexinteraction of a variety of pro-inflammatory or anti-inflammatorystimulators or mediators, which are synthesized and released by tissue.Known species of pro-inflammatory or anti-inflammatory stimulators ormediators include, but are by no means limited to, cytokines, nitricoxide, thromboxanes, leukotrienes, platelet-activating factor,prostaglandins, kinins, complement factors, superantigens, monokines,chemokines, interferons, free radicals, proteases, arachidonic acidmetabolites, prostacyclins, beta endorphins, myocardial depressantfactors, anandamide, 2-arachidonoylglycerol, tetrahydrobiopterin, andchemicals including histamine, bradykinin, and serotonin. The discoveryof new (i.e., previously unrecognized) species of pro-inflammatory oranti-inflammatory stimulators or mediators occurs almost daily.

The nature and intensity of inflammatory responses differ, depending onthe site which has been invaded, and on the character of theInflammatory Agent(s), and the interaction of pro-inflammatory oranti-inflammatory stimulators or mediators involved.

The inflammatory response, when regulated and localized, is beneficial.However, if not regulated and generalized, the inflammatory response cancause significant tissue injury and even death.

For example, cytokines are a class of proteins produced by macrophages,monocytes, and lymphocytes in response to viral or bacterial infection,as well as in response to T cell stimulation during an immune response.Cytokines are normally present in very low concentrations in the bloodor tissues.

The structures and activities of cytokines have been the subject of manystudies. It has become apparent that cytokines possess a wide spectrumof immunological and non-immunological activities. It is also apparentthat cytokines affect diverse physiologic functions, such as cellgrowth, differentiation, homeostasis and pathological physiology. It isclear that cytokines have multiple biological activities and interactwith more than one cell type. Cytokines are also known to be capable ofstimulating their own synthesis, as well as the production of othercytokines from a variety of cell types. This phenomenon is called the“cytokine cascade.”

Cytokine cascades are associated with systemic changes arising frominfection and tissue injury and, in this context, they serve a myriad ofbiological functions. For example, various cytokines, categorized as theinterleukins (IL), interferons (IF), and tumor necrosis factor (TNF),are produced during immune and inflammatory responses. These cytokinesbeneficially control various aspects of these responses. In thissituation, the cytokine cascade mediates normal host defense responses,cell regulation, and cell differentiation.

However, it has been observed that the function of cytokine productioncan become disordered. This can lead to the presence of larger thannormal concentrations of cytokines. When the cytokine cascade becomesdisordered, there can be a rapid extension and amplification of theintended localized host response in such a way that only one or a fewinitiating stimuli trigger the eventual release and participation ofscores of host mediators. Although a number of features of the hostresponse assist in fighting off invasion, an overly robust or poorlymodulated endogenous response can rapidly accelerate to produce otherprofound alterations in host homeostasis at the cellular, tissue, andsystemic levels. As a result, cytokine expression in a region of thebody where tissues or organs are legitimately subject to bacterialinfection or an immune response challenge, can, when disordered, lead tounwanted destruction of healthy tissue elsewhere in the body. Largerthan normal concentrations of certain cytokines can cause disease andother deleterious health effects, some of which can be lethal.

For example, a disordered cytokine cascade that leads to the increasedpresence of the cytokines IL-1 and TNF can, alone or in combination,cause a state in animals clinically identical to “septic” shock. It isrecognized that septic shock arises due to the individual, combined, andconcerted effects of a large number of cytokines. It is a conditioninflicting more than 450,000 Americans every year. Cytokine-inducedseptic shock can be brought about by infection by a variety ofmicroorganisms, including not only bacteria but also viruses, fungi, andparasites. Septic shock can also be initiated by host response toinvasion in general, such as by cancer or as a result of major surgeryor trauma. Septic shock is a potentially lethal cytokine-mediatedclinical complication against which there is no generally effectivetherapeutic approach.

One of the best studied examples of cytokine-induced septic shock is thecase of infection by gram-negative bacteria. It is believed that theappearance of bacterial endotoxins, such as lipopolysaccharide (LPS), inthe host bloodstream leads to the endogenous production of a variety ofhost factors that directly and indirectly mediate the toxicity of LPS.These host-derived mediators include many now well-recognizedinflammatory cytokines, as well as endocrine hormones, in addition to anumber of other endogenous factors such as leukotrienes and plateletactivating factor. Among the interacting factors that together comprisethe cytokine cascade, the cytokine TNF alpha is believed to be the mostimportant identified to date. During the ensuing cytokine cascade, themediators that appear early in the invaded host are thought to triggerthe release of later appearing factors. Many of the cytokine mediatorsnot only exert direct functions at the targeted tissues, but also atother local and remote tissues, where subsequent responses to othermediators produced during the cascade occur, and so on. The result, ifunchecked, can be a multifaceted pathological condition, which ischaracterized most prominently by deleterious hemodynamic changes andcoagulopathy leading to multiple organ failure and, often, to death.

Multiple attempts have been made and still many others are currentlyunderway to block specific mediators of this response. These attemptshave been relatively unsuccessful. Therapy aimed at single mediatorscannot effectively attenuate the entire response. Furthermore, it is theduration rather than the intensity of inflammation that correlates bestwith outcome, in that the longer the duration of over-expression ofproinflammatory cytokines the higher the mortality. Systemicinflammation results in organ injury which results in the prolongationof the inflammatory response and thus, more organ injury.

Less lethal but just as profound physiologic effects can occur as aresult of abnormal production of certain cytokines, without the presenceof exogenous bacterial toxins. As one example, cytokine TNF-alpha hasbeen found to be an anti-tumor cytokine. As a result, TNF-alpha has beenexpected to be useful as an antitumor agent. However, it has beendiscovered that TNF-alpha is identical with cachectin, which is acachexia-inducing factor. The disordered production of TNF-alpha hasalso been correlated with, not only septic shock, but the incidence ofrheumatoid arthritis, adult respiratory distress syndrome (ARDS), theseverity of viral hepatitis, myocardial ischemia, and the inhibition ofmyocardial contraction. Also, TNF has recently been shown to be involvedin initiating the expression of human immunodeficiency virus in humancells that carry latent virus, which could be a contributing factor inthe expression of latent AIDS virus in certain individuals. Furthermore,a correlation between the TNF level in the blood and blood pressure hasalso been observed. As TNF levels increase, blood pressure decreases,which can lead to serious complications such as kidney failure.

It has also been observed that TNF-alpha also has an activity ofstimulating production of other types of cytokines, such as IL-1, etc.It is known that the cytokine IL-1 is an important agent for inducingand transmitting the systemic biological response against infection andinflammation. IL-1 induces the usual, desirable responses observed ininflammation in general, such as fever, increase of leukocytes,activation of lymphocytes, induction of biosynthesis of acute phaseprotein in liver. It also known that this cytokine has a strongantitumor activity.

However, when IL-1 is produced in abnormally larger amounts, it maycontribute to the severity of chronic inflammatory diseases, such asrheumatoid arthritis. Thus, the abnormal activation of various cytokinessuch as the interleukins (IL) and tumor necrosis factor (TNF) isbelieved responsible for the tissue damage and pain that occurs invarious inflammatory conditions like rheumatoid arthritis. In rheumatoidarthritis, levels of TNF, IL-1, IL-6 and IL-8 increase dramatically andcan be detected in the synovial fluid. The cytokine cascade induced byexpression of these cytokines results in depressed lipoproteinmetabolism as well as bone and cartilage destruction.

As another example, the cytokine IL-6 plays an important role inantibody production in B cells. The cytokine IL-6 also is an importantfactor in body systems, e.g., the hematopoietic system, nervous system,and the liver, as well as in immune system. For example, IL-6 iseffective for inducing proliferation and differentiation of T cells,inducing the production of protein at acute phase by acting on hepaticcells, and promoting the growth of cells in bone marrow.

However, it has also been observed that there is a correlation betweenthe abnormal secretion of IL-6 and various disease states, e.g.,autoimmune diseases, such as hypergammaglobulinemia, chronic articularrheumatism, and systemic lupus erythematosus; the abnormal state ofpolyclonal B cells, as well as in the development of the abnormal stateof monoclonal B cells such as myeloma cells; Castleman's diseaseaccompanied with tumor of the lymph nodes, for which the cause isunknown; primary glomerular nephritis; and the growth of mesangialcells.

As yet another example, in bacterial infections, cytokines such as IL-8act as a signal that attracts white blood cells such as neutrophils tothe region of cytokine expression. In general, the release of enzymesand superoxide anions by neutrophils is essential for destroying theinfecting bacteria. However, if cytokine expression causes neutrophilsto invade, for example, the lungs, release of neutrophil enzymes andsuperoxide anion can result in the development of adult respiratorydistress syndrome (ARDS), which can be lethal.

Despite their diverse and myriad functions, all cytokines share onecommon feature. They are all within a narrow size and molecular weightrange of 8 to 28 kilodaltons. This size characteristic is extremelyimportant for the clearance of cytokines from the blood. In this range,cytokines are effectively cleared by the liver and also the kidney,which clears all proteins below 50 kilodaltons in size. An imbalancebetween cytokine production and cytokine removal can cause damage to theliver and kidney.

In disease states where the kidney has failed—which is often the case inseptic shock—hemodialysis or hemofiltration membranes are used assubstitutes for the glomerular membrane of the kidney. However,artificial membranes are severely limited in their ability to clearcytokines from the blood due to their inadequate porosity. In fact, thepredominant mechanism by which these membranes remove cytokines inclinical practice is not filtration, but rather nonspecific surfaceadsorption (J. Am Soc Nephrol 1999 April; 10(4): 846-53, Cytokineremoval during continuous hemofiltration in septic patients, De Vriese AS, Colardyn F A, Philippe J J, Vanholder R C, De Sutter J H, Lameire NH). Typically these membranes have 0.5 to 2 square meters of surfacearea available for adsorption that becomes saturated within the first 30to 90 minutes of treatment (Biomaterials September 1999; 20(17):1621-34,Adsorption of low molecular weight proteins to hemodialysis membranes:experimental results and simulations, Valette P, Thomas M, Dejardin P).

It is therefore clear that pro-inflammatory or anti-inflammatorystimulators or mediators, such as cytokines but by no means limited tocytokines, have the potential for both desirable physiologic results andundesirable physiologic results, depending upon the robustness andmodulation of a particular inflammatory response. There is a need forstraightforward and biocompatible devices, systems, and methods thatserve to reduce or otherwise modulate levels of pro-inflammatory oranti-inflammatory stimulators or mediators in instances where abnormallevels of or unregulated or excessive interaction among such materialsexist or can be expected to arise.

SUMMARY OF THE INVENTION

A detrimental inflammatory response, such as may occur, e.g., in thecontinuum from early sepsis to septic shock, or ischemia reperfusion,allograft rejection, chemical/biologic warfare casualties, hastraditionally been viewed as a condition in which the local inflammatoryresponse has become generalized and uncontrolled. Immune effector cells,especially neutrophils, possess potent cytotoxic capacity and whenunchecked, this response can cause significant tissue injury.

However, while this traditional view is true, these intense inflammatoryresponse conditions may also be viewed as a syndrome of immunesuppression. Immune effector cells become dysfunctional and are nolonger capable of normal immune surveillance. Such a condition resultsin increased susceptibility to recurrent infection, prolongedinflammation and continued tissue injury. This condition can be referredto as “immuno-paralysis” and can be easily demonstrated. When eitherintact septic animals or whole blood taken from septic patients isexposed to an inflammatory stimulus (e.g. endotoxin) the normal hostresponse is severely inhibited.

From this perspective, therapy aimed at reducing an inflammatoryresponse by targeting removal of some of the pro-inflammatory stimulusmay not restore normal immune responsiveness and thus, may not improveoutcome. Instead, a more desirable immune modulating strategy is to usea biocompatible adsorption medium to selectively adsorb a broaderspectrum of pro-inflammatory or anti-inflammatory stimulators ormediators, which may include but is not neccesarily limited tocytokines, and to thereby restore immunologic stability, rather thanindiscriminately inhibiting or stimulating one or another component.Such a strategy counters the immunologic instability of sepsis and otherintense inflammatory response conditions by reducing the number, andthus the activity, of a wide array of both pro- and anti-inflammatorymolecules. Such a strategy would “auto-regulate” itself, such that asone component of the response increased so too would the effect on thatcomponent. Finally, the desirable strategy might well be limited in itseffect to the circulating pool of mediators rather than influencing thetissue levels where their activity may be beneficial.

The invention provides devices, systems, and methods for reducing levelsof cytokines or other species of pro-inflammatory or anti-inflammatorystimulators or mediators in the blood, desirably whole blood, or bloodproducts, or physiologic fluids in situations where abnormal levels ofor unregulated or excessive interaction among such stimulators ormediators occur, or during events that do induce or have the potentialfor inducing abnormal production of or unregulated or excessiveinteraction among such stimulators or mediators. The devices, systems,and methods serve to prevent, control, reduce, modulate, or alleviatethe severity of many physiologic conditions and disease states that areassociated with abnormal levels of or unregulated or excessiveinteraction among pro-inflammatory or anti-inflammatory stimulators ormediators.

One aspect of the invention provides devices, systems, and methods forremoving cytokines or other species of pro-inflammatory oranti-inflammatory stimulators or mediators from the blood, desirablywhole blood, which are of use in acute situations where abnormal levelsor unregulated or excessive interaction among such stimulators ormediators are present in individuals experiencing infection, orindividuals experiencing an immune response. In such situations, thedevices, systems and methods serve to modulate the inflammatory responseby removing at least some of these stimulators or mediators from bloodcirculation, even as such stimulators or mediators are being produced bythe individual to fight off the infection or invasion. This aspect ofthe invention serves to prevent an overly robust endogenous response,such as occurs, e.g., during septic shock. The devices, systems, andmethods can be used alone or in combination with other forms oftreatment targeted to the treatment of the bacterial infection and/orimmune response.

Another aspect of the invention provides devices, systems, and methodsfor removing cytokines or other species of pro-inflammatory oranti-inflammatory stimulators or mediators from the blood, desirablywhole blood, which are of use in situations where abnormal levels of orunregulated or excessive interaction among such stimulators or mediatorsare or may be present, or which involve events that do induce or havethe potential for inducing abnormal production of or unregulated orexcessive interaction among such stimulators or mediators in certain “atrisk” individuals undergoing or about to undergo surgery, e.g., fortreatment of burns or cardiac conditions; or for organ transplantationor reconstructive surgery, or other episodes involvingischemia-reperfusion injury. Other like situations, where abnormallevels of or unregulated or excessive interaction among such stimulatorsor mediators are or may be present, or which involve events that doinduce or have the potential for inducing abnormal production of orunregulated or excessive interaction among such stimulators ormediators, include certain “at risk” individuals who have experiencedtrauma, such as burns, or “the crush syndrome.” In such situations, thedevices, systems, and methods serve to reduce the population of suchstimulators or mediators by removing at least some of such stimulatorsor mediators from the blood circulation. This aspect of the inventionalso serves to modulate the inflammatory response by removing at leastsome pro-inflammatory or anti-inflammatory stimulators or mediators fromthe blood circulation, even as such stimulators or mediators are beingproduced by the individual in response to the surgery or trauma. Thisaspect of the invention serves to prevent an overly robust endogenousresponse, to prevent, e.g., septic shock or other conditions that mayoccur.

Another aspect of the invention provides devices, systems, and methodsfor removing cytokines or other species of pro-inflammatory oranti-inflammatory stimulators or mediators from the blood, desirablywhole blood, which are of use in situations where abnormal cytokinelevels are present in certain “at risk” individuals, whose chronicdisease states are caused by or otherwise correlate with increasedinflammatory activity. Such disease states include, e.g., rheumatoidarthritis; or lung disease such as emphysema or asthma; or pulmonaryfailure; or adult respiratory distress syndrome (ARDS); viral hepatitis;or myocardial ischemia; or autoimmune disease; AIDS; or as a result ofaccidental or intentional exposure to biological or chemical agents,such as anthrax. In such situations, the devices, systems, and methodsserve to reduce the population of cytokines or or other species ofpro-inflammatory or anti-inflammatory stimulators or mediators byremoving such stimulators or mediators from the blood circulation. Thisaspect of the invention serves to treat a given disease condition bylessening the abnormal population of cytokines or other species ofpro-inflammatory or anti-inflammatory stimulators or mediators, which isknown or suspected of contributing to severity of the disease condition.The devices, systems and methods can be used alone or in combinationwith other treatment modalities for the disease condition.

Another aspect of the invention provides devices, systems, and methodsfor removing cytokines or other species of pro-inflammatory oranti-inflammatory stimulators or mediators from the blood, which are ofuse in other events that do induce or have the potential for inducingproduction of such stimulators or mediators due to extracorporeal bloodprocessing, handling, or storage. These events can lead to an incidentalor “obligatory” activation of the immune system due to subjecting theblood to extracorporeal treatment, pumping, or storage, e.g., forcentrifugal or membrane blood separation; or for hemodialysis orhemofiltration; or for oxygenation. This obligatory activation of theimmune system can activate production of cytokines or or other speciesof pro-inflammatory or anti-inflammatory stimulators or mediators in theblood as it undergoes extracorporeal treatment, handling, or storage.The increased presence of cytokines or other species of pro-inflammatoryor anti-inflammatory stimulators or mediators in the treated, handled,or stored blood or blood product can, upon re-infusion, generate anincidental inflammatory response in the recipient's system, or at leastcan contribute to an incidental abnormal level of cytokines or otherspecies of pro-inflammatory or anti-inflammatory stimulators ormediators in the recipient. In such events, the devices, systems andmethods serve to reduce the population of cytokines or other species ofpro-inflammatory or anti-inflammatory stimulators or mediators byremoving such stimulators or mediators from the treated, handled, orstored blood or blood product. This aspect of the invention serves toprevent incidental inflammatory response conditions or disease states asa result of otherwise beneficial blood treatment, handling or storage,by lessening the population of cytokines or other species ofpro-inflammatory or anti-inflammatory stimulators or mediators presentin the re-infused blood or blood product.

The devices, systems, and methods that embody features of the inventionalso make it possible to restore a normal balance betweenpro-inflammatory stimulators or mediators and anti-inflammatorystimulators or mediators. For example, during a cytokine cascade,pro-inflammatory cytokines are typically generated in larger numbers inproportion to anti-inflammatory cytokines. In situations where abnormalcytokine levels exist, the removal of cytokines according to theinvention will tend to remove more pro-inflammatory cytokines thananti-inflammatory cytokines, and thereby aid in maintaining a morenormal balance between the two.

Another aspect of the invention provides devices, systems, and methodsfor removing cytokines or other species of pro-inflammatory oranti-inflammatory stimulators or mediators from physiologic fluids. Forexample, spent peritoneal dialysis solution can carry cytokines or otherspecies of pro-inflammatory or anti-inflammatory stimulators ormediators. Systems and methods exist for regenerating spent peritonealdialysis solution withdrawn from a patient, by removing waste and uremictoxins from the spent solution, as well as introducing electrolytes andbuffering materials into the spent solution. In this way, freshperitoneal dialysis solution can be recreated, obviating the need forbagged replacement solutions. In such situations, the devices, systems,and methods that embody this aspect of the invention remove cytokines orother species of pro-inflammatory or anti-inflammatory stimulators ormediators from the peritoneal dialysis solution, before, during, orafter solution regeneration. This aspect of the invention serves toprevent incidental inflammatory response conditions or disease states asa result of exchange of spent peritoneal dialysis solution withregenerated peritoneal dialysis solution.

As another example, organs harvested for transplantation, e.g., kidney,liver, or heart, are typically stored for period of time in a suitablepreservation solution until transplantation takes place. Storage of theorgan in preservation solution can lead to the generation of cytokinesor other species of pro-inflammatory or anti-inflammatory stimulators ormediators, which accumulate in the preservation solution. In suchsituations, the devices, systems, and methods that embody this aspect ofthe invention remove cytokines or other species of pro-inflammatory oranti-inflammatory stimulators or mediators from the preservationsolution during organ storage and/or before transplantation of the organoccurs. In this way, the invention serves to prevent or at leastameliorate inflammatory response conditions or disease states as aresult of organ transplantation.

As yet another example, body fluids that are removed from and thenrecycled back to the body during a given treatment modality can carrycytokines or other species of pro-inflammatory or anti-inflammatorystimulators or mediators, or cytokines or other species ofpro-inflammatory or anti-inflammatory stimulators or mediators can begenerated as a result of such treatment modalities. Treatment systemsand methods exist for removing and recycling such fluids, e.g.,lymphatic fluid, synovial fluid, spinal fluid, or cerebrospinal fluid.The devices, systems, and methods that embody this aspect of theinvention can be used in association with such treatment modalities, toremove cytokines or other species of pro-inflammatory oranti-inflammatory stimulators or mediators from the body fluids before,during, or after primary treatment.

In preferred embodiments, the devices, systems, and methods removecytokines or other species of pro-inflammatory or anti-inflammatorystimulators or mediators from the blood by selective adsorption.Desirably, the selective adsorption medium is characterized by abiocompatibility index that reflects a negligible production ofcytokines or other species of pro-inflammatory or anti-inflammatorystimulators or mediators in the blood as a result of exposure to themedium. Thus, the adsorption medium, which beneficially serves to removecytokines or other species of pro-inflammatory or anti-inflammatorystimulators or mediators from the blood, does not itself produce anoffsetting result of generating additional cytokines or other species ofpro-inflammatory or anti-inflammatory stimulators or mediators.

Other features and advantages of the inventions are set forth in thefollowing specification and attached drawings.

BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1 is a schematic view of a system for removing cytokines or otherspecies of pro-inflammatory or anti-inflammatory stimulators ormediators from the blood in acute or chronic or other “at risk”situations;

FIG. 2 is a schematic view of a system for removing cytokines or otherspecies of pro-inflammatory or anti-inflammatory stimulators ormediators from the blood during an extracorporeal blood processingprocedure, such as blood separation, dialysis, hemofiltration, orextracorporeal oxygenation;

FIG. 3 is a side section view of a unitary, extracorporeal devicecontaining an adsorption medium for removing cytokines or other speciesof pro-inflammatory or anti-inflammatory stimulators or mediators fromthe blood;

FIG. 4A is a side view of an exchangeable device that can be coupled toa conventional intravenous blood access catheter for the purpose ofremoving cytokines or other species of pro-inflammatory oranti-inflammatory stimulators or mediators from the blood;

FIG. 4B is a side view of the exchangeable device shown in FIG. 4A afterbeing coupled to a conventional intravenous blood access catheter forthe purpose of removing cytokines or other species of pro-inflammatoryor anti-inflammatory stimulators or mediators from the blood;

FIG. 5 is a side section view of an intravenous catheter having a wallthat is impregnated with an adsorption material that removes cytokinesor other species of pro-inflammatory or anti-inflammatory stimulators ormediators from the blood;

FIG. 6 is a side section view of an intravenous catheter having anintegrally formed chamber containing an adsorption medium that removescytokines or other species of pro-inflammatory or anti-inflammatorystimulators or mediators from the blood;

FIG. 7 is a side view of an indwelling catheter having an in-line devicethat contains an adsorption medium for removing cytokines or otherspecies of pro-inflammatory or anti-inflammatory stimulators ormediators from the blood, making possible an ambulatory treatmentregime;

FIG. 8 is a side section view of a composite treatment module whichintegrates a device for removing cytokines or other species ofpro-inflammatory or anti-inflammatory stimulators or mediators from theblood with a blood processor, the removal device being shown connectedby intermediate tubing downstream from the blood processor;

FIG. 9 is a side section view of a composite treatment module whichintegrates a device for removing cytokines or other species ofpro-inflammatory or anti-inflammatory stimulators or mediators from theblood with a blood processor, the removal device being shown connectedby intermediate tubing upstream from the blood processor;

FIG. 10A is a side section view of a composite treatment module whichintegrates a device for removing cytokines or other species ofpro-inflammatory or anti-inflammatory stimulators or mediators from theblood with a blood processor, the removal device and the blood processorcomprising separate units adapted to be joined together for use;

FIG. 10B is the composite treatment module shown in FIG. 10B after beingjoined together for use;

FIG. 11 is a side section view of a composite treatment module whichintegrates a device for removing cytokines or other species ofpro-inflammatory or anti-inflammatory stimulators or mediators from theblood with a blood processor, the module comprising a common housingcompartmentalized into two chambers, one chamber containing the bloodprocessing component and the other chamber containing an adsorptionmedium for removing cytokines or other species of pro-inflammatory oranti-inflammatory stimulators or mediators from the blood beingprocessed;

FIG. 12 is a side section view of an adsorption particle that can beused in association with the systems shown in FIGS. 1 and 2 forselectively adsorbing cytokines or other species of pro-inflammatory oranti-inflammatory stimulators or mediators from the blood;

FIG. 13 is a side section view of a device that is usable in associationwith the systems shown in FIGS. 1 and 2 for removing both cytokines orother species of pro-inflammatory or anti-inflammatory stimulators ormediators and other targeted proteins or toxins from the blood;

FIG. 14 is a schematic view of a system for removing cytokines or otherspecies of pro-inflammatory or anti-inflammatory stimulators ormediators from a physiologic fluid, which takes the form of regeneratedperitioneal dialysis solution;

FIG. 15 is a schematic view of a system for removing cytokines or otherspecies of pro-inflammatory or anti-inflammatory stimulators ormediators from a physiologic fluid, which takes the form of preservationsolution for an organ awaiting transplantation;

FIG. 16 is a schematic diagram of a test system that is used tocharacterize the biocompatibility index of a given adsorbant medium;

FIG. 17 is a graph plotting the cytokine response in the blood of asepsis animal model as a result of treatment using a biocompatibleadsorbant medium;

FIGS. 18A, 18B, and 18C are graphs showing the variations in blood cellcounts for red blood cells, white blood cells, and platelets,respectively, during passage of 25 ml of the blood through a treatmentdevice containing an adsorbant medium useful for removing cytokines fromthe blood;

FIG. 19 is a graph showing the variations in PMN elastase concentrations(indicative of leukocyte activation) during passage of 25 ml of theblood through a treatment device containing an adsorbant medium usefulfor removing cytokines from the blood;

FIG. 20 is a graph showing the variations in LDH concentrations(indicative of hemolysis) during passage of 25 ml of the blood through atreatment device containing an adsorbant medium useful for removingcytokines from the blood;

FIG. 21 is a graph showing the variations in C3a-desArg concentrations(indicative of complement activation) during passage of 25 ml of theblood through a treatment device containing an adsorbant medium usefulfor removing cytokines from the blood;

FIG. 22 is a graph showing the variations in TAT concentrations(indicative of coagulation) during passage of 25 ml of the blood througha treatment device containing an adsorbant medium useful for removingcytokines from the blood; and

FIG. 23 is a chart summarizes the results of hemocompatibility testingconducted by Bosch et al of a polyacrylate gel adsorbant material (forthe selective adsorption of low-density lipoproteins), based uponcontact with blood that was anticoagulated either only with heparin orwith a mixture of heparin and citrate.

The invention may be embodied in several forms without departing fromits spirit or essential characteristics. The scope of the invention isdefined in the appended claims, rather than in the specific descriptionpreceding them. All embodiments that fall within the meaning and rangeof equivalency of the claims are therefore intended to be embraced bythe claims.

DESCRIPTION OF THE PREFERRED EMBODIMENTS

I. Systems and Methods for Removing Cytokines From the Blood

Cytokines and other species of pro-inflammatory or anti-inflammatorystimulators or mediators are low molecular weight proteins that arepresent in the blood. They are typically produced by the body inresponse to viral or bacterial infection and in response an immuneresponse. Cytokines are also known to be capable of stimulating theirown synthesis, as well as the production of other cytokines from avariety of cell types. Cytokines are normally present in very lowconcentrations in a tissue, but, due to an over-robust and unmodulatedcytokine cascade or other causes, cytokines can be present in abnormalconcentrations. In abnormal concentrations, cytokines can cause diseaseor septic shock.

As used in this Specification, the term “cytokine” as used herein ismeant any secreted polypeptide that affects the functions of othercells, and is a molecule which modulates interactions between cells inthe immune or inflammatory response. Cytokines are soluble protein andpeptide humoral regulators. Type-1 cytokines are produced by Type-1helper cells, e.g. IL2 , IFN-gamma , IL12 and TNF-beta, and Type-2cytokines are produced by Type-2 helper cells, e.g. IL4, IL5, IL6, IL10,and IL13. These may be pro-inflammatory or anti-inflammatory,chemotactic, paracrine, endocrine, juxtacrine, autocrine, andretrocrine. They also function as growth factors and apoptosis factors,involved in inflammation, septic shock, the systemic inflammatoryresponse syndrome (SIRS), acute phase reactions, wound healing andneuroimmune networks. Others include IFN-alpha, -beta, -gamma, -omega,IL2-9, GCSF, MCSF, GMCSF, PGDF, IL-1-alpha, -beta, TNF-alpha, FGF, IL8,IP10, PF4, GRO, 9E3 and recombinant cytokines, muteins, and proteinmimetics. Cytokines also comprise B-cell differentiation factors (BCDF),Bcell growth factors (BCGF), mitogenic cytokines, chemotactic cytokines(chemokines), colony stimulating factor (CSF), angiogenesis factors,t-cell replacing factor (TRF), heparin binding growth factor (HBGF),substance p (tachykinin), and kinins.

A. Acute or “At Risk” Conditions

FIG. 1 generically shows a system 10 for removing cytokines 12 or otherspecies of pro-inflammatory or anti-inflammatory stimulators ormediators (which are generally identified by circled C's in FIG. 1) fromthe blood 14, and desirably from whole blood. In the illustratedembodiment, the blood 14 emanates from a blood source 16. In theembodiment shown in FIG. 1, it is contemplated that the blood source 16comprises the circulatory system of an individual.

In FIG. 1, it is also contemplated that the cytokines 12 or otherspecies of pro-inflammatory or anti-inflammatory stimulators ormediators exist in the blood in abnormal levels, or at least thepotential exists that the individual's levels of cytokines or otherspecies of pro-inflammatory or anti-inflammatory stimulators ormediators may become abnormal, i.e., reach levels above normalphysiologic levels, or otherwise create an unregulated or excessiveinflammatory response interaction. Accordingly, as shown in FIG. 1, thesystem 10 includes a device 18 through which the blood 14 is circulatedfrom the source 16 for the purpose of removing at least a portion of thepopulation of cytokines 12 or other species of pro-inflammatory oranti-inflammatory stimulators or mediators carried in the blood 14. Theremoval of cytokines 12 or other species of pro-inflammatory oranti-inflammatory stimulators or mediators from the blood 14 serves tocontrol, reduce, or alleviate the severity of many physiologicconditions and disease states that are associated with abnormal cytokinelevels or an unregulated or excessive inflammatory response. As shown inFIG. 1, the cytokine-depleted blood 20 is returned to the individualblood source 16.

The cytokines 12 or other species of pro-inflammatory oranti-inflammatory stimulators or mediators may be present or pose thepotential to exist in the blood 14 in abnormal levels for variousreasons. For example, the individual may be in an acute condition,experiencing infection or an immune response. In this situation,cytokines 12 or other species of pro-inflammatory or anti-inflammatorystimulators or mediators are being generated by the individual to fightthe infection or invasion. The concurrent removal of cytokines 12 orother species of pro-inflammatory or anti-inflammatory stimulators ormediators by the device 18 modulates the inflammatory response, e.g., toprevent the onset of a condition on a continuum from sepsis to septicshock or damage to tissue elsewhere in the body. Alternatively, theindividual may be experiencing a condition on a continuum from sepsis toseptic shock. In this situation, the concurrent removal of cytokines 12or other species of pro-inflammatory or anti-inflammatory stimulators ormediators by the device 18 modulates the inflammatory response toterminate the deleterious hemodynamic changes and coagulopathyoccasioned by septic shock, to prevent organ failure and death. Ineither situation, one prevention and the other treatment, the removal ofcytokines 12 or other species of pro-inflammatory or anti-inflammatorystimulators or mediators by the device 18 aims to prevent an overlyrobust and possible lethal endogenous response.

The device 18 can be used alone or in combination with other forms oftreatment targeted to the treatment of the bacterial infection and/orimmune response and/or septic shock. Examples of other forms oftreatment that can be used in combination with the device 18 includeantibiotics, antimicrobial agents, antifungal agents, antiviral agents,and specific compounds such as activated protein-C.

In another embodiment, the cytokines 12 or other species ofpro-inflammatory or anti-inflammatory stimulators or mediators may bepresent in abnormal levels because the individual possesses an “at risk”acute or chronic disease state, which is caused by or otherwisecorrelate with increased physiologic cytokine activity or an unregulatedinflammatory response. Such disease states include, e.g., rheumatoidarthritis; or lung disease such as emphysema or asthma; or pulmonaryfailure; or adult respiratory distress syndrome (ARDS); viral hepatitis;or myocardial ischemia; or autoimmune disease; AIDS; or as a result ofexposure to biological or chemical agents, such as anthrax. The removalof cytokines 12 or other species of pro-inflammatory oranti-inflammatory stimulators or mediators by the device 18 reduces thepopulation of cytokines or other species of pro-inflammatory oranti-inflammatory stimulators or mediators to treat the severity of thedisease condition. The treatment of the individual using the system 10can be under acute conditions (due to the presence of severe symptoms).The treatment using the system 10 can also be under chronic conditions,as a part of scheduled, periodic treatment of the disease condition.

In either situation, the device 18 can be used alone or in combinationwith other treatment modalities beneficial for the disease condition.Examples of other forms of treatment that can be used in combinationwith the device 18 include antibiotics, antimicrobial agents, antifungalagents, antiviral agents, and specific compounds such as activatedprotein-C.

In another embodiment, the cytokines 12 or other species ofpro-inflammatory or anti-inflammatory stimulators or mediators may bepresent in abnormal levels, or may potentially rise to abnormal levels,because the individual is “at risk” due to present or contemplatedsurgery, e.g., for treatment of burns or cardiac conditions; or fororgan transplantation or reconstructive surgery, or other episodesinvolving ischemia- reperfusion injury. Alternatively, the individualcan be “at risk” because of trauma, such as burns, or “the crushsyndrome,” which may or may not require corrective surgery. In suchsituations, cytokines 12 or other species of pro-inflammatory oranti-inflammatory stimulators or mediators have likely already beengenerated by the individual due to injury and trauma to the body, andresulting corrective surgery is likely to maintain or even increasegeneration of cytokines or other species of pro-inflammatory oranti-inflammatory stimulators or mediators. The removal of cytokines 12or other species of pro-inflammatory or anti-inflammatory stimulators ormediators by the device 18, after the trauma and either before surgery,or during surgery, or after surgery, or a combination thereof, reducesthe population of cytokines or other species of pro-inflammatory oranti-inflammatory stimulators or mediators, to modulate the inflammatoryresponse. The removal of cytokines 12 or other species ofpro-inflammatory or anti-inflammatory stimulators or mediators by thedevice 18 aims to prevent an overly robust and possible lethalendogenous response, to prevent, e.g., septic shock or other unregulatedor excessive inflammatory response conditions that may occur. Thetreatment using the system 10 can occur under acute conditions (i.e., asan adjunct to the surgical procedure or other treatment of the trauma),and/or under chronic conditions, as a part of a scheduled rehabilitationprogram following the trauma or surgery.

In either situation, the device 18 can be used alone or in combinationwith other treatment modalities beneficial for the injury and surgicalprocedure. Examples of other forms of treatment that can be used incombination with the device 18 include antibiotics, antimicrobialagents, antifungal agents, antiviral agents, and specific compounds suchas activated protein-C.

B. Extracorporeal Blood Processing

FIG. 2 show a blood processing system 20 that removes cytokines 12 orother species of pro-inflammatory or anti-inflammatory stimulators ormediators from the blood 14 as it undergoes extracorporeal processing.In use, the system 20 is intended to convey the blood from a bloodsource 22 (typically, the circulatory system of a donor or patient) toan extracorpreal blood processing assembly 24. After processing, all ora portion of the blood is either returned to the circulatory system ofthe individual donor or patient, or retained for storage and subsequenttransfusion to the same donor or patient, or to another recipient, or acombination thereof.

Typically, the functional components of the blood processing assembly 24are a blood inlet line 26, a blood processor 28, and a blood outlet line27. The blood from the donor or patient is conveyed by the blood inletline 26 to the processor 28 for the desired processing. Afterprocessing, the blood is convey from the processor 28 by the bloodoutlet line 27. The system 20 may continuously or intermittently conveythe blood to and from the blood processing assembly 24, typically usingone or more peristaltic pumps (designated P in FIG. 2).

Depending upon the objectives of the processing, the blood outlet line27 can be coupled directly to the donor or patient, so that theprocessed blood is returned directly to that individual. In otherprocessing schemes, all or a portion of the processed blood is retainedfor storage and not returned to the donor or patient. In thisarrangement, the blood outlet line 27 also communicates with a bloodstorage container 32.

The blood processing assembly 24 can be constructed in various ways andperform different processing functions.

1. Blood Separation

The blood processing assembly 24 can serve to separate whole blood intoplasma and cellular blood components (i.e., blood products), typically,red blood cells and platelets. In this arrangement, the blood processingassembly 24 can comprise a centrifuge or a membrane that separates wholeblood into its components. Depending upon the objectives of the device,all or some of the components are collected for storage and latertransfusion. The components that are not collected are typicallyreturned to the blood donor.

For example, in a process called plasmapheresis, plasma can be collectedin an extracorpeal circuit for later fractionation to harvesttherapeutic plasma proteins, e.g., Factor VIII. The remaining cellularcomponents (red blood cells and platelets, along with the leukocytes)are returned to the blood donor.

Or, in a process called plasma exchange, plasma can be collected in anextracorpreal circuit. The plasma is discarded, and the cellularcomponents (red blood cells, leukocytes, and platelets) are returned tothe blood donor, along with a plasma-replacement fluid. Alternatively,the plasma itself can be treated by immunoadsorption, to removeundesired materials—e.g., antibodies—which is then returned with thecellular components to the individual.

As another example, in a process called plateletpheresis, the blood iscirculated through an extracorpreal path through a centrifuge, whichcentrifugally separates and collects concentrated platelets for latertransfusion. The remaining cellular components and plasma are returnedto the donor. Alternatively, a volume of red blood cells or plasma, orboth, can be retained for storage and later transfusion to recipientsundergoing blood component therapy.

There are many other types of blood cell harvesting procedures inaddition to plateletpheresis, where a targeted blood cell is collected,e.g., leukopheresis. There are also many other types of blood processingprocedures in general, such as photopheresis (for inactivation of viralpathogens) or hypothermia, which circulate blood in extracorporeal pathsto achieve desired therapeutic or diagnostic objectives.

The preceding examples process the blood on-line, that is, while thedonor remains coupled to the system. In another arrangement, calledmanual collection, a unit of whole blood is drawn into a plastic bloodcollection bag, to which one or more plastic satellite bags areintegrally connected. These arrangements of integrally connected bagsare called multiple blood bag systems. After the unit of whole blood isdrawn, the donor is disconnected. The whole blood is then subjected tooff-line centrifugation while in the blood collection bag. Thecentrifugation separates the whole blood into layers of red blood cellsand plasma, with an intermediate layer of leukocytes. The plasma can beeither rich in platelets or poor in platelets, depending upon thecentrifugal forces applied. The plasma component is transferred into asatellite bags, leaving the red blood cells (and leukocytes) behind inthe blood collection bag. If rich in platelets, the plasma component canbe further centrifugally separated in the satellite bag to obtainconcentrated platelets. The components are stored in the individualplastic bags for later transfusion to recipients undergoing bloodcomponent therapy.

2. Hemodialysis or Hemofiltration

The blood processing assembly 24 can also carry out processes, calledhemodialysis or hemofiltration, which emulate normal kidney activitiesfor an individual whose renal function is impaired or lacking.

During hemodialysis, the blood from an individual is conveyed in anextracorporeal path along one side of a membrane. A dialysate iscirculated on the other side of the membrane and forms a concentrationdifferential across the membrane. Liquid and uremic toxins carried inthe blood are drawn by the concentration differential across themembrane and out of the blood.

During hemofiltration, the blood from an individual is conveyed in anextracorporeal path along a semipermeable membrane, across which apressure difference (called transmembrane pressure) exists. The pores ofthe membrane have a molecular weight cut-off that can pass liquid anduremic toxins carried in the blood.

In both hemodialysis and hemofiltration, the membrane pores do not passformed cellular blood elements and plasma proteins. These components areretained and returned to the individual with the toxin-depleted blood,along with a replacement fluid. The replacement fluid restores, at leastpartially, a normal physiologic fluid and electrolytic balance to theblood. Hemodialysis and hemofiltration can be carried out as individualprocesses, or in combination.

A form of hemodialysis is also used to treat individuals suffering fromjaundice caused by inadequate liver function or liver failure. In thisindication, the blood carries abnormal levels of bilirubin, a breakdownproduct of hemoglobin normally removed by the liver. The blood is passedalong one side of a dialysis membrane. Healthy liver cells are locatedon the opposite side of the membrane. The healthy liver cells removebilirubin from the processed blood. In this treatment, the blood ispassed before undergoing dialysis through an adsorption device(typically contained activated charcoal) to remove certain bloodmaterials that are lethal to liver cells.

3. Oxygenation (Cardiopulmonary Bypass)

The blood processing assembly 24 can alternatively carry out a processcalled oxygenation. Oxygenation is carried out during cardiopulmonarybypass, during which the blood is circulated outside the heart and lungswhile heart surgery occurs. During oxygenation, the blood conveyed froman individual is transported in an extracorporeal path along a membraneacross which a oxygen concentration differential exists. Oxygen from theopposite side of the membrane is transported into the blood on theopposite side of the membrane, to emulate lung function.

4. Removal of Cytokines or Other Species of Pro-Inflammatory orAnti-Inflammatory Stimulators or Mediators

Extracorporeal processing of the blood in the system 20 may trigger anincidental or “obligatory” activation of the components of the immunesystem carried by the blood. The sources of this incidental activationcan include exposure to biomaterials in the inlet and return lines 26and 28 or in the blood processing assembly 24 itself. External pumpingof the blood can also trigger an incidental immune response. Thecentrifugal forces or shear forces developed by passage along a membranecan also trigger an incidental immune response.

The incidental activation of the immune system occasioned during bloodprocessing can lead to the incidental generation of cytokines or otherspecies of pro-inflammatory or anti-inflammatory stimulators ormediators. These cytokines or other species of pro-inflammatory oranti-inflammatory stimulators or mediators, to the extent that they areincidentally produced as a result of blood processing, will betransported by the blood that is returned to the donor or patient duringprocessing, or by stored blood delivered to a recipient duringtransfusion. Entering the circulatory system of the donor or otherrecipient, these incidental cytokines or other species ofpro-inflammatory or anti-inflammatory stimulators or mediators can serveto raise the levels of cytokines or other species of pro-inflammatory oranti-inflammatory stimulators or mediators in the donor or otherrecipient, and could lead to the generation of further cascades orinflammatory responses, during which further cytokines or other speciesof pro-inflammatory or anti-inflammatory stimulators or mediators andadditional by-products of immune system activation are produced. Thus,processes that provide beneficial results in one respect can lead toincidental, potentially adverse results in another respect.

The blood processing system 20 therefore includes a device 30 thatremoves cytokines or other species of pro-inflammatory oranti-inflammatory stimulators or mediators from the processed blood.

In on-line blood processing systems—e.g., those systems in which thecirculatory system of the donor or patient remains coupled to theprocessor 24 during processing—the device 30 can be coupled in-lineeither upstream or downstream of the processor 24 (in FIG. 2, the device30 is shown positioned in the return line 28 for purposes ofillustration). In this arrangement, cytokines or other species ofpro-inflammatory or anti-inflammatory stimulators or mediators areremoved during circulation of the blood through the extracorporealcircuit, thereby leading to reduced levels of cytokines or other speciesof pro-inflammatory or anti-inflammatory stimulators or mediators in theblood returned to the donor or patient.

In off-line blood processing systems—e.g., where the blood is processedafter disconnecting the donor from the collection system—or in a systemthat collects a blood component for later transfusion to a recipient (asFIG. 2 shows)—it is desirable to place the device 30 either upstream ofthe blood component storage bag (as shown in phantom lines in FIG. 2)(so cytokines or other species of pro-inflammatory or anti-inflammatorystimulators or mediators are removed after blood processing and beforestorage of the blood component) or in a transfusion set coupled to thesatellite blood component storage bag (so that cytokines or otherspecies of pro-inflammatory or anti-inflammatory stimulators ormediators are removed during the act of transfusion of the processedblood component).

The device 30 serves to reduce the population of cytokines or otherspecies of pro-inflammatory or anti-inflammatory stimulators ormediators by removing cytokines or other species of pro-inflammatory oranti-inflammatory stimulators or mediators from the treated, handled, orstored blood. The device 30 thereby serves to prevent incidentalcytokine-induced or inflammatory response conditions or disease statesas a result of otherwise beneficial blood treatment, handling orstorage, by lessening the population of cytokines or other species ofpro-inflammatory or anti-inflammatory stimulators or mediators presentin the returned or re-infused blood. The removal by the device 30 ofcytokines or other species of pro-inflammatory or anti-inflammatorystimulators or mediators generated as a result of extracorporeal bloodprocessing aims to maintain a status quo condition in the immune systemof the individual undergoing blood processing or the recipient of storedblood.

II. Devices for Removing Cytokines or Other Species of Pro-Inflammatoryor Anti-Inflammatory Stimulators or Mediators From the Blood

Cytokines and other species of pro-inflammatory or anti-inflammatorystimulators or mediators are low molecular weight, electrically neutralproteins, ranging in size from about 8000 to about 28,000 daltons.Cytokines or other species of pro-inflammatory or anti-inflammatorystimulators or mediators can be removed from the blood by variousmechanism, e.g. by selective adsorption, or by ion exchange, or bynon-specific adsorption to dialysis membranes. The devices 18 or 30 forremoving cytokines or other species of pro-inflammatory oranti-inflammatory stimulators or mediators from the blood can thereforebe variously constructed, depending upon the removal mechanism selected.

In the illustrated embodiment, selective removal by adsorption is theselected mechanism.

A. Unitary Extracorporeal Devices

Either device 18 or 30 can comprise a stand-alone, or unitary,extracorporeal component that can be coupled in-line to blood tubing attime of use.

In this arrangement (see FIG. 3), either device 18 or 30 desirablyincludes in its most basic form a housing 32. The housing 32 contains amedium 34 that removes cytokines or other species of pro-inflammatory oranti-inflammatory stimulators or mediators by adsorption.

The housing 32 includes an inlet 33 for conveying the blood into thehousing 32 for contact with the adsorption medium 34. The housing 32also includes an outlet 36 for conveying the blood from the housingafter contact with the adsorption medium 34, during which all or aportion of the cytokines or other species of pro-inflammatory oranti-inflammatory stimulators or mediators present are removed.

Desired characteristics of the adsorption medium 34 will be described ingreater detail later.

The transport of the blood through the adsorption medium 34 in thehousing 32 can be accomplished in various ways, depending in large partupon the environment in which the device 18 or 30 is used. In the acuteor chronic applications described, which involve use of the device 18,an external pump can be used to convey the blood through the housing 32to remove cytokines or other species of pro-inflammatory oranti-inflammatory stimulators or mediators. Alternatively, blood tubingconnected to the inlet 33 of the housing 32 can be coupled via asuitable blood access to an artery, while blood tubing connected to theoutlet 36 of the housing 32 can be coupled by a suitable blood access toa vein, thereby using physiologic blood pressure to convey the bloodthrough the housing 32 to remove cytokines or other species ofpro-inflammatory or anti-inflammatory stimulators or mediators.

When used in association with a blood processing system, which involvesuse of the device 30, an external pump (identified as P in FIG. 2) istypically present to convey the blood through the blood processingassembly 24. In this arrangement, the external pump P that serves theblood processing assembly can concurrently provide the pressure toconvey the blood through the housing 32 to remove cytokines or otherspecies of pro-inflammatory or anti-inflammatory stimulators ormediators.

In an alternative embodiment shown in FIGS. 4A and 4B, the housing 32can be configured to comprise an exchangeable component 38 that can bereleasably coupled to a conventional intravenous blood access catheter40, e.g., of the type widely used in intensive care units. Theexchangeable component 38 provides particular ease of use in eitheracute or chronic indications, as above described, as individuals in suchcircumstances are typically already fitted with intravenous blood accesscatheters for other purposes. However, the exchangeable components 38would also provide ease of use in the setting of extracorporeal bloodprocessing, as the intravenous blood tubing comprising the blood inletline 26 or blood outlet line 27 serving the processor 28 could be readymodified to include fittings to accommodate quick exchange of thecomponent 38.

In this arrangement, the inlet 33 and 36 of the exchangeable component38 and the catheter 40 (or inlet and outlet lines 26 and 27) wouldinclude, e.g., convention mating luer fittings 42, to enable quickattachment and removal in-line in the intravenous blood access catheter40 or intravenous blood lines 26/27 serving the processor 28, as FIGS.4A and 4B demonstrate.

In another alternative embodiment shown in FIG. 5, all or a portion ofthe wall of an intravenous catheter 44 can be impregnated with theadsorption medium 34. In this arrangement, transport of the bloodthrough the catheter 44 exposes the blood to the medium 34 for theremoval of cytokines or other species of pro-inflammatory oranti-inflammatory stimulators or mediators. Alternatively (as shown inFIG. 6), an intravenous catheter 46 can include an integrally formedchamber 48 in which the adsorption medium 34 is housed. Thus, transportof the blood through the catheter 44 exposes the blood to the medium 34for the removal of cytokines or other species of pro-inflammatory oranti-inflammatory stimulators or mediators. In the embodiments shown inFIGS. 5 and 6, the device 18 or 30 forms an integrated part of the bloodtransport path, so that a separate housing 32 per se is not required tocontain the adsorption medium 34.

B. Ambulatory Applications

As FIG. 7 shows, either device 18 or 30 can comprise a component 50 thatis intended to be coupled to an indwelling catheter 52, that issurgically fitted to the individual undergoing treatment. The catheter52 is surgically attached to the circulatory system of the individual,e.g., between an artery and a vein, to form a loop through which theblood continuously circulates. In this arrangement, the component 50carries the adsorption medium 34 that serves to remove cytokines orother species of pro-inflammatory or anti-inflammatory stimulators ormediators from the individual's blood traversing the catheter 52. As apart of an indwelling blood circulation loop, the component 50 removescytokines or other species of pro-inflammatory or anti-inflammatorystimulators or mediators continuously on a daily basis, as theindividual ambulates and carries on life's activities outside of atreatment facility.

The component 50 can be configured to be an external or internalexchangeable device that can be releasable coupled to the indwellingcatheter 52, e.g., by use of luer fittings 42, in the manner generallyshown in FIGS. 4A and 4B. Alternatively, the wall of the indwellingcatheter 52 can itself be impregnated with the adsorption medium 34, asgenerally shown in FIG. 5.

The component 50, in association with an indwelling catheter 52, makespossible a continuous, ambulatory treatment to remove cytokines or otherspecies of pro-inflammatory or anti-inflammatory stimulators ormediators. This treatment modality would have particular application forthose “at risk” individuals whose disease states are caused by orotherwise correlate with chronic, increased physiologic cytokineactivity or other unregulated inflammatory response condition. Thecomponent 50 provides a new form of ambulatory treatment for, e.g.,rheumatoid arthritis; or lung disease such as emphysema or asthma; oradult respiratory distress syndrome (ARDS); or autoimmune disease; orAIDS. The component 50 serves to maintain a reduced population ofcytokines or other species of pro-inflammatory or anti-inflammatorystimulators or mediators, by continuously removing cytokines or otherspecies of pro-inflammatory or anti-inflammatory stimulators ormediators from the blood circulation. The component 50 can be used aloneor in combination with other treatment modalities for the diseasecondition.

C. Integrated Composite Devices

FIGS. 8 and 9 show an absorption device 30 of a type shown in FIG. 3,integrally coupled by intermediate tubing 43 to a blood processor 28.Together, the device 30, processor 28, and linking tubing 43 form acomposite blood treatment module 54 that is supplied to a user as anintegrated unit.

The composite module 54 can be arranged so that the absorption device 30is integrally coupled in a downstream flow direction to the bloodprocessor 28 (as FIG. 8 shows), or, alternatively arranged, in anupstream flow direction to the blood processor 28 (as FIG. 9 shows). Inyet another arrangement, the adsorption device 30 can be placed bothupstream and downstream of the blood processor 28.

The module 54 can perform different blood processing functions inassociation with a blood adsorption function, e.g., to remove cytokinesor other species of pro-inflammatory or anti-inflammatory stimulators ormediators, depending upon the operational capabilities of the bloodprocessor 28. The processor 28 can be configured to perform diversefunctions, e.g., hemodialysis, or hemofiltration, or membrane separationof plasma from whole blood, or blood filtering (e.g., to removeleukocytes), or ionic exchange, etc., or combinations thereof.

As FIGS. 10A and 10B show, the adsorption device 30 can be moreintimately attached to the blood processor 28 to form the module 54without use of intermediate tubing 43. In this arrangement (see FIG.10A), both the adsorption device 30 and processor 28 are manufactured asseparate units. The adsorption device 30 and processor 28 are configuredwith, e.g., a tubular male fitting 56 on the device 30 that mates with afemale fitting 58 in the processor 28. The fittings 56 and 58 couple thedevice 30 and the processor 28 together in fluid flow communication, asFIG. 10B shows.

Of course, the mating configuration of the fittings 56 and 58 can bereversed, so that the device 30 includes a female fitting 58 and theprocessor 28 includes the male fitting 56. Furthermore, other attachmentconfigurations, e.g., screw fit, keyed fittings, etc., can be used.Mating stabilization struts 60 may also be provided to further lock thedevice 30 and processor 28 together.

By manufacturing the adsorption device 30 and separator 28 separately,and then joining them together to form an integrated module 54,different sterilization processes may be used. For example, the device30 and adsorption medium 34 may be sterilized by a first sterilizationprocess, e.g., hot water or steam or external irradiation, whereas theprocessor 28 may be sterilized by a second, different sterilizationprocess, e.g., EtO sterilization. This modular arrangement therebyaccommodates the choice of biomaterials for the adsorption medium 34 andthe functional component of the processor 28 having different physicalproperties best suited for their particular functional objections, andnot constrained by similar sterilization requirements. The arrangementshown in FIGS. 8 and 9 also accommodates different sterilizationtechniques prior to joining the device 30 and processor 28 with thetubing 43.

As with the embodiments shown in FIGS. 8 and 9, the fittings 56 and 58can configured to join the device 30 in an upstream flow direction tothe blood processor 28, or (as FIG. 10B shows) in a downstream flowdirection to the blood processor 28, or at both upstream and downstreamends of the blood processor 28.

The device 30 may be integrally coupled to the processor 28 duringmanufacturing, and be supplied to the customer as an integrated module54 (as FIG. 10B shows). Alternatively, the device 30 and processor 28may be supplied separately to the customer (in the manner shown in FIG.10A), who is instructed to join the adsorption device 30 to theprocessor 28 by plugging the fittings 56 and 58 together at time of use.

As FIG. 11 shows, the adsorption device 30 can be even more intimatelyassociated with the blood processor by placing the processor 28 anddevice 30 within the confines of a single housing 62. The single housing62 has an inlet port 68 and an outlet port 70. In this arrangement, aninterior partition wall 72 in the housing 62 compartmentalizes thehousing 62 into a first compartment 64 (which communicates with theinlet port 68) and a second compartment 66 (which communicates with theoutlet port 70). One or more openings 74 in the interior wall 72 openflow communication between the first and second compartments 64 and 66.

Each compartment 64 and 66 can contain either the functional componentof the processor 28 or the adsorption medium 34. In the embodiment shownin FIG. 11, the functional component of the processor 28 is contained inthe first compartment 64, and the adsorption medium 34 is contained inthe second compartment 66. Of course, the arrangement of the materialscontained in the compartments 64 and 66 can be reversed. The housing canalso be partitioned to place the adsorption medium 34 at both the inletand outlet sides of the blood processor 28, sandwiching the functionalcomponent of the blood processor 28 between it.

This arrangement requires the selection of materials for the processor28 and adsorption medium 34 that accommodate the same sterilizationprocess, e.g., hot water sterilization.

It should be appreciated that the various composite structures 54 justdiscussed, which join an adsorption device 30 with a blood processor 28,are not limited to a particular adsorption function for the adsorptiondevice 30. That is, while the adsorption device 30 has be earlierdescribed in this application the context of the removal of cytokines orother species of pro-inflammatory or anti-inflammatory stimulators ormediators, the adsorption device 30 can, in association with theprocessor 28, carry out other functions as well. For example, when theprocessor 28 takes the form of a hemodialyzer, the adsorption device 30can serve the function of selectively adsorbing middle molecular weightproteins (e.g., beta-2 macroglobulin) that conventional hemodialysismembrane do not efficiently remove.

D. Adsorption Medium

The adsorption medium 34 can be variously constructed. In theillustrated embodiment (see, e.g., FIG. 3), the adsorption medium 34desirable includes a group of porous polymeric particles 76, which areformed to selectively retain cytokines or other species ofpro-inflammatory or anti-inflammatory stimulators or mediators. Takinginto account the physical proportions of cytokines or other species ofpro-inflammatory or anti-inflammatory stimulators or mediators, thepolymeric particles 76 of the medium 34 are predominantly mesoporous,with a pore size ranging from 2 to 70 nm, and preferably from 5 to 50nm.

As FIG. 12 best shows, each polymer particle 76 desirably possesses aporous hydrophobic core 78. The pores are sized to provide close contactbetween the cytokines or other species of pro-inflammatory oranti-inflammatory stimulators or mediators and the hydrophobic surfaceof the pores.

The surface of the hydrophobic particles 76 can be modified to provide ahydrophilic coating 80, which imparts a high degree of biocompatibilitywith the human organism, and, in particular, the blood. Thisbiocompatibility can be expressed in terms of a biocompatible index, aswill be described in greater detail later. The hydrophilic coating 80 isdesirably thin and permeable so as to allow penetration of cytokines orother species of pro-inflammatory or anti-inflammatory stimulators ormediators to the hydrophobic porous core 78 of the particles 76.

The hydrophobic cores 78 of the particles 76 can be composed, forexample, of crosslinked polymeric materials prepared by polymerizationor copolymerization of the following monomers: styrene, ethylstyrene,α-methylstyrene, divinylbenzene, di isopropenyl benzene,trivinylbenzene, alkyl methacrylate as methyl methacrylate, butylmethacrylate. The hydrophilic biocompatible coating 80 of the particles76 can be composed for example of the following materials:polyvinylpyrrolidone, polyhydroxyethyl methacrylate,carboxymethylcellulose, polyurethane.

In a device of the type shown in FIG. 3, the particles 76 are sized,taking into account the size of the device, to obtain a desired flowrate through the device. As an example, given a device size of 400 ml,the particles 76 are sized greater than 300 μm in diameter to present aneffective surface area to the blood of about 500 m²/gram of adsorptionmedium 34 used.

Particles 76 having the characteristics described also selectivelyadsorb superantigens. Superantigens are low molecular weight proteinsthat are toxic. Superantigens are produced by organisms and are strongactivators of the immune system and cytokine production. The presence ofsuperantigens can therefore also contribute to increased levels ofcytokines or other species of pro-inflammatory or anti-inflammatorystimulators or mediators. The concurrent removal by the particles ofboth cytokines or other species of pro-inflammatory or anti-inflammatorystimulators or mediators and superantigens enhances the overalltherapeutic function of the adsorption medium 34.

Representative Adsorption Medium EXAMPLE 1

In one representative embodiment, the adsorption medium 34 can includeparticles or beads formed from hypercrosslinked polystyrene-type resins.The surface of the beads is desirably modified to prevent absorption oflarge proteins and platelet and to minimize activation of bloodcomplement system, without affecting noticeably accessibility of aninner absorption space of the beads for small and middle-sizedmolecules. The particles or beads can comprise, e.g.,styrene-divinylbenzene copolymers subjected to an extensive crosslinkingin a swollen state with bifunctional crosslinking agents, such asmonochlorodimethyl ether or p-xylylene dichloride. Alternatively, theparticles or beads can comprise styrene-divinylbenzene copolymerssubjected to chloromethylation and post-crosslinking. Alternatively, thematerial can comprise a porous hydrophobic acrylic polymer or amesoporous ethylstyrene-divinylbenzene copolymer.

The surface modification can be accomplished is various ways, e.g., (i)by depositing on the surface of the particles or beads high molecularweight poly(N-trifluoroalkoxy) phosphazene, by treating the beads with asolution of phosphazene in an organic solvent and evaporating thesolvent; or (ii) electrostatically binding of heparin from its aqueoussolution onto the beads whose chloromethyl groups have been substitutedby amino functions through a reaction with an amine, such as 2-ethanolamine; (iii) substituting chloromethyl groups on the surface of thebeads with 2-ethanol amine ligands and covalently binding heparin to theligands via a material such as a glutare dialdehyde and hexamethylenediisocyanate moiety, and coupling groups consisting of excessive pendantaldehyde groups and isocyanate groups with L-aspartic acid; or (iv)substituting chloromethyl groups with a material such as 2-ethanol amineand ethylene glycol ligands, activating the ligands with a material suchas glutare dialdehyde and hexamethylene diisocynate, and covalentlybinding hydrophilic polyethylene glycol chains; or (v) covalentlybinding hydrophilic polyethylene glycol chains through reacting ofsodium alcoholates of the latter with polystyrene chloromethyl groups;or (vi) covalently binding hydrophilic chains of chitosan throughreacting of amino groups of the latter with polystyrene chloromethylgroups; or (vii) substituting chloromethyl groups with ligands such as2-ethanol amine ligands or ethylene glycol ligands, activating theligands with phosphorus oxychloride, and covalently binding hydrophilicmoieties such as choline, serine and 2-ethanol amine.

Further details regarding the composition of particles or beads of thistype can be found in U.S. Pat. No. 5,904,663, which is incorporatedherein by reference.

Representative Adsorption Medium EXAMPLE 2

In another representative embodiment, the adsorption medium 34 caninclude particles or beads formed from a porous hydrophobicdivinylbenzene copolymer with comonomers selected from the group ofstyrene, ethylstyrene, acrylonitrile, and buthyl methacrylate. Suchparticles or beads initially have surface exposed vinyl groups, whichare chemically modified to impart improved biocompatibility, so as toform different surface exposed functional groups, such as polymers of2-hydroxyethyl methacrylate, N-vinylpyrrolidine, N-vinylcaprolactame, orN-acrylamide. The surface exposed functional groups can be products ofoxidation of the vinyl groups to expoxy groups and subsequent additionof polar compounds selected from the group of water, ethylene glycol,primary or secondary amines, and 2-hydroxethyl-amine. Alternatively, thesurface exposed functional groups can be the products of oxidation ofthe vinyl groups to epoxy groups, the subsequent addition of primary orsecondary amines or 2-hydroxyethylamine, and the deposit ofhigh-molecular-weight poly(trifluoroethoxy)phosphazene.

Further details regarding the composition of particles or beads of thistype can be found in U.S. Pat. No. 6,114,466, which is incorporatedherein by reference.

Representative Adsorption Medium EXAMPLE 3

In another representative embodiment, the adsorption medium 34 caninclude particles or beads formed by polymerization of aromatic divinylcompounds, such as p- or m-divinylbenzene or mixtures thereof, or theircopolymerization with aromatic monovinyl compounds, such as styrene,methylstyrene, ethylvinylbenzene and vinylbenzylchloride, in thepresence of porogens or mixtures of porogens with properties close tothose of θ-solvents. The porogens can comprise, e.g., cyclohexane,cyclohexanone and other θ-solvents for polystyrene. Alternatively, theporogens can comprise θ-solvents composed of mixtures of a good solventfor polystyrene, such as toluene, benzene, ethylene dichloride,propylene dichloride, tetrachloroethene, dioxane and methylenedichloride, and a non-solvent for polystyrene, such as aliphatichydrocarbons, aliphatic alcohols and aliphatic acids.

Such hypercrosslinked polymeric adsorbents exhibit a combination ofmicropores, mesopores and macropores. The adsorbents may further befunctionalized to enhance their biocompatibility.

Further details regarding the composition of particles or beads of thistype can be found in U.S. patent application Ser. No. 09/143,407, filedAug. 28, 1998, entitled “Hypercrosslinked Polymeric Material forPurification of Physiological Liquids of Organism, a Method forProducing the Material,” which is incorporated herein by reference.

1. Biocompatibility Index

Desirably, the adsorption medium 34 is characterized by abiocompatibility index that indicates a physiologically negligibleproduction of cytokines or other species of pro-inflammatory oranti-inflammatory stimulators or mediators in the blood as a result toexposure to the medium. Thus, the adsorption medium 34, which beneficialserves to remove cytokines or other species of pro-inflammatory oranti-inflammatory stimulators or mediators from the blood, does notitself produce an offsetting result of generating additional cytokinesor other species of pro-inflammatory or anti-inflammatory stimulators ormediators.

The biocompatibility index can be expressed as a dimensionless, numericquantity, which reflects the degree to which a prescribed battery ofblood characteristics change as a result of contact between the bloodand the adsorption medium.

The prescribed battery of blood characteristics that thebiocompatibility index encompasses rely upon several selected bloodindicators, which quantify, based upon contact between the blood and agiven adsorption medium, (i) the degree to which the numbers of cellularblood components (red blood cells, white blood cells, and platelets) arediminished; (ii) the degree to which leukocytes are activated; (iii) thedegree to which complement activation occurs; (iv) the degree to whichhemolysis occurs; and (v) the degree to which clot formation is induced.

Indicator (i) is ascertained by Coulter Counter for red blood cells,white blood cells, and platelets (this indicator this comprises threeindividual indicators).

Indicator (ii) is ascertained by measuring polymorphonuclear leukocyteelastase (PMN Elastase) concentrations using standard laboratorytechniques (e.g., PMN Elastase, Merck Immunoassay, Merk KgaA, Darmstadt,Germany).

Indicator (iii) is ascertained by measuring anaphylatoxin C3a-desArgconcentrations using standard laboratory techniques (e.g., Elisa, ProgenBiotechnik GmbH, Heidelberg, Germany).

Indicator (iv) is ascertained by determining the concentrations ofLactate dehydrogenase (LDH) by standard methods of clinical chemistry.

Indicator (v) is ascertained by measuring the concentrations ofthrombin-antithrombin-complex (TAT) using standard laboratory techniques(e.g., Enzygnost-TAT micro Elisa, Dade Behring Marburg GmbH, Marburg,Germany).

There are therefore a total of seven indicators within the battery ofindicators for the Biocompatibility Index: (1) White Blood Cell Count;(2) Red Blood Cell Count; (3) Platelet Count; (4) PMN ElastaseConcentration; (5) LDH Concentration; (6) C3a-desArg Concentration; and(7) TAT Concentration. These indicators are listed in Table 1, below.

In deriving the biocompatibility index, the technician selects a housingfor the media that is made of an acceptable biocompatible material thatpossesses a biocompatibility comparable to conventional medical gradeplastics (e.g., polyvinylchloride, polyurethane, polyester, etc) orglass. The technician characterizes the blood according to the batteryof indicators after passing the blood through the housing in an emptycondition, i.e., a housing that contains no absorption medium.

The technician uses heparin to anticoagulate the blood in a finalconcentration of 1.0 IU heparin/ml blood. Other types of anticoagulant,such as nafamosat, may be used. However, citrate anticoagulant is not beto used, alone or in combination with the prescribed amount of heparinin deriving the biocompatibility-index, because the presence of citratewill mask changes in thrombogenicity and complement activation that mayarise due to contact with the medium, thereby leading to false results.

FIG. 23 summarizes the results of hemocompatibility testing conducted byBosch et al of a polyacrylate gel adsorbant material (for the selectiveadsorption of low-density lipoproteins), based upon contact with bloodthat was anticoagulated either only with heparin or with a mixture ofheparin and citrate (Bosch et al, Artif Organ 17(7) 640-52 1993). FIG.23 demonstrates that, with respect to the thrombogenicity and complementactivation indicators—PMN Elastase (indicating the degree to whichleukocytes are activated); thrombin-antithrombin-complex TAT (indicatingthe degree to which clot formation is induced); and anaphylatoxinC3a-desArg (indicating the degree to which complement activationoccurs)—each indicator level reads high (denoting thrombogenicity andcomplement activation) when only heparin anticoagulant is used. Themixture of citrate with heparin masks the actual indicator levels in asignificant way. FIG. 23 shows that, by binding calcium (an importantco-factor in many hemocompatibility reactions), the presence of citratelowers the indicator levels, so that they no longer reflect the actualchanges in thrombogenicity and complement activation that arise due tocontact with a given medium.

The forgoing protocol provides the background or baseline sample,against which the magnitude of changes due to the presence of a givenadsorption medium within the housing can be ascertained and scored.

In deriving the biocompatibility index, the technician alsocharacterizes the blood according to the battery of indicators afterpassage through the selected housing that contains the absorptionmedium.

As before, the technician uses heparin to anticoagulate the blood in afinal concentration of 1.0 IU heparin/ml blood. For the reasons statedabove, citrate anticoagulant is not be to used in deriving thebiocompatibility index, alone or in combination with the prescribedamount of heparin.

In carrying out the steps just described, the technician assembles atest system 300 as shown in FIG. 16. The test system 300 comprises twoparallel channels 302 and 304 connected by a y-connector 306 to a bloodline 308. A housing 310 and 312 is coupled in each channel, respectively302 and 304. The housing 310 is empty (i.e., free of adsorption medium),and the housing 312 contains the adsorption medium 314. The blood line308 can be coupled, e.g., to the antecubital vein of a healthyvolunteer. The access system desirably allows for continuousheparinization at the tip of the inserted cannula or needle to avoidsystemic heparinzation.

Peristaltic pumps P1 and P2 in the channels 302 and 304 (or a single,double tube peristaltic pump) convey the blood through the housings 310and 314. An infusion pump P3 meters heperin, to achieve a final heparinconcentration of 1.0 IU/ml.

The pumps P1, P2, and P3 are started simultaneously. On-line bloodperfusion of the two channels 302 and 304 is maintained through eachhousing 310 and 312. The speeds of the pumps P1 and P2 are adjusted to10 mL/min through each housing 310 and 312. Blood samples are collectedat the outlet of each channel 302 and 304 after 5, 10, 15, and 25minutes of perfusion directly into specially prepared polypropylenevials V stored on ice. The blood samples are analyzed for the selectedindicators immediately. Blood counts are corrected for hemodilution dueto the addition of heparin.

The cell count indicators are corrected by the following formula:X_(corr)=X times (hct_(pre)/hct_(t), where X_(corr) is the correctedparameter, X is the measured value of the parameter at time point t,hct_(pre) is the hematocit pre value (t=0), and hct_(t) is thehematocrit at time point t.

The plasma indicators for PMN Elastase Concentration, LDH Concentration,C3a-desArg Concentration, and TAT Concentration are corrected by thefollowing formula: X_(corr)=X times (1-hct_(t)/1-hct_(pre), whereX_(corr) is the corrected plasma parameter, X is the measured value ofthe plasma parameter at time point t, hct_(pre) is the hematocit prevalue (t=0), and hct_(t) is the hematocrit at time point t.

The technician reviews the assembled indicators to ascertain, for eachindicator, the maximum difference between the indicator values over 25ml of blood flow of the blood passed through the housing 310 (withoutthe medium—baseline) and the blood passed through the housing 312containing the medium 314. For each indicator, the technician expressesthe maximum change as a percentage, relative to the baseline value.

The technician then scores the percentage change for each indicator as adimensionless numeric quantity 1, 2, or 3, depending upon the magnitudeof the percentage change, in accordance with Table 1. In Table 1, apercentage change equal to or less than a prescribed minimum for a givenindicator is scored as a 1, signifying a most desirable degree ofbiocompatibility. In Table 1, a percentage change greater than aprescribed maximum for a given indicator is scored as a 3, signifying aleast desirable degree of biocompatibility. In Table 1, a percentagechange between the prescribed minimum and the prescribed maximum for agiven indicator is scored as a 2, signifying an acceptable degree ofbiocompatibility, albeit not the most desired.

TABLE 1 The Biocompatibility Index Score Table Numeric Scores 1 2 3(Signifying (Signifying (Signifying Most an a Least Desired AcceptableDesired Blood Degree of Degree of Degree of Indicator Biocompatibility)Biocompatibility) Biocompatibility) Loss of White Maximum MaximumMaximum Blood Cells Difference Difference Difference Between BetweenBetween Baseline Baseline and Baseline and and Medium Medium (25 ml)Medium (25 ml) (25 ml) >15% >20% ≦15% ≦20% Loss of Red Maximum MaximumMaximum Blood Cells Difference Difference Difference Between BetweenBetween Baseline Baseline and Baseline and and Medium Medium (25 ml)Medium (25 ml) (25 ml) >15% >20% ≦15% ≦20% Loss of Maximum MaximumMaximum Platelets Difference Difference Difference Between BetweenBetween Baseline Baseline and Baseline and and Medium Medium (25 ml)Medium (25 ml) (25 ml) >15% >20% ≦15% ≦20% PMN Elastase Maximum MaximumMaximum Concentration Difference Difference Difference Between BetweenBetween Baseline Baseline and Baseline and and Medium Medium (25 ml)Medium (25 ml) (25 ml) >15% >20% ≦15% ≦20% LDH Maximum Maximum MaximumConcentration Difference Difference Difference Between Between BetweenBaseline Baseline and Baseline and and Medium Medium (25 ml) Medium (25ml) (25 ml) >15% >20% ≦15% ≦20% C3a-desArg Maximum Maximum MaximumConcentration Difference Difference Difference Between Between BetweenBaseline Baseline and Baseline and and Medium Medium (25 ml) Medium (25ml) (25 ml) >20% >25% ≦20% ≦25% TAT Maximum Maximum MaximumConcentration Difference Difference Difference Between Between BetweenBaseline Baseline and Baseline and and Medium Medium (25 ml) Medium (25ml) (25 ml) >15% >20% ≦15% ≦20%

After scoring each indicator with a numeric quantity of 1, 2, or 3, thetechnician adds the numeric quantities scored for all the indicators toobtain a total. The total constitutes the biocompatibility index for thegiven adsorption medium.

The Biocompatibility Index for a given material is a reliable indicatorof blood compatibility. There is a strong between the value of theBiocompatibility Index, derived in the manner just described, and theability of given material to selectively remove targeted proteins fromthe blood without significant destruction of cellular components andhemolysis and without significant clot formation (i.e., lowthrombogenicity). Materials characterized by a Biocompatibility Indexequal to or less than 14, and, most desirably, by a Biocompatible Indexnot greater than 7, contact the blood with no significant loss of bloodcells, no significant hemolysis, no significant activation of luekocytesor monocytes, and, at most, only very mild complement activation, evenwith the use of heparin as the sole anticoagulant. Because suchmaterials are not likely to induce the generation of cytokines or otherspecies of pro-inflammatory or anti-inflammatory stimulators ormediators, they are well suited for use to remove cytokines or otherspecies of pro-inflammatory or anti-inflammatory stimulators ormediators from the blood, blood products, or physiologic fluids.

On the other hand, materials characterized by a Biocompatibility Indexgreater than 14, contact the blood with adverse effects in terms ofsignificant blood cell loss, or significant hemolysis, or significantleukocyte activation, or significant compliment activation, orsignificant combinations thereof. Such materials are therefore likely toinduce the generation of cytokines or other species of pro-inflammatoryor anti-inflammatory stimulators or mediators and are not acceptable foruse to remove cytokines or other species of pro-inflammatory oranti-inflammatory stimulators or mediators.

E. Multiple Functionality

As previously discussed, the devices, systems, and methods are directedto the removal of cytokines or other species of pro-inflammatory oranti-inflammatory stimulators or mediators to reduce levels of suchagents in the blood in situations where abnormal levels of such agentsoccur, or during events that do induce or have the potential forinducing abnormal production of cytokines or other species ofpro-inflammatory or anti-inflammatory stimulators or mediators. In thisway, the devices, systems, and methods serve to control, reduce, oralleviate the severity of many physiologic conditions and disease statesthat are associated with abnormal levels of cytokines or other speciesof pro-inflammatory or anti-inflammatory stimulators or mediators.

It should be appreciated that the devices, systems, and methods can beadapted to perform other functions in tandem with removal of cytokinesor other species of pro-inflammatory or anti-inflammatory stimulators ormediators as well.

FIG. 13 shows a device 82 that is usable in association with the systemsand methods previously discussed to provide adsorption of both cytokinesor other species of pro-inflammatory or anti-inflammatory stimulators ormediators and other material or materials from the blood. The device 82includes a first compartment 84, which contains the adsorption medium34, previously described, to remove cytokines or other species ofpro-inflammatory or anti-inflammatory stimulators or mediators. Thedevice 82 includes a second compartment 86, which contains a differentmedium 88, which can comprise an adsorption medium or an ion exchangemedium, to remove another type of material from the blood. A partition90 in the device 82 (e.g., made of a mesh material to accommodate fluidflow) separates the first compartment 84 from the second compartment 86.

In use, the blood is conveyed into the device 82 through an inlet 92.The blood passes in succession through the adsorption medium 34 and thedifferent, second medium 88. The blood exits the device 82 through anoutlet 94. During passage, cytokines or other species ofpro-inflammatory or anti-inflammatory stimulators or mediators areremoved from the blood by the adsorption medium 34 and the othermaterial is removed from the blood by the different, second medium 88.The order of passage through the mediums 34 and 88 can be reversed.

The adsorption medium 88 can be variously constructed depending upon thematerial intended to be removed.

1. Removal of LPS EndoToxin

For example, the adsorption medium 88 can be constructed to remove LPSendotoxin, which is released into the blood of an individual sufferingfrom a gram-negative bacterial infection. In the blood, LPS endotoxincoalesce into vesicles ranging in size from 300,000 to 1,000,000daltons. Phosphoryl groups contained within the LPS endotoxin give it anoverall negative charge at physiologic pH. The release of LPS endotoxininto the blood can cause fever, low blood pressure, and organ failure.

As previously discussed, the presence of LPS endotoxin also stimulatesthe secretion of cytokines or other species of pro-inflammatory oranti-inflammatory stimulators or mediators. The presence of LPSendotoxin can therefore also contribute to increased levels of cytokinesor other species of pro-inflammatory or anti-inflammatory stimulators ormediators, and even to the onset of a septic shock episode.

In the illustrated embodiment (see FIG. 13), the adsorption medium 88includes a group of polymer particles 96 comprising hydrophobic porouscore to which LPS endotoxin binds. To provide a reliable interactionbetween the endotoxin and the polymer core, the polymer particles havepores of a corresponding large size. For example, the size of the porescan be within the range of 20 to 150 nm, and preferably between 30 and100 nm. The polymeric particles 96 are thus predominantly macroporous.

The polymer for the core of the particles 96 can be selected from thesame group of materials as the polymer for the core 78 of the particles76 of the adsorption medium 34, as before described.

Like the particles 76 of the first adsorption medium 34, the particles96 of the adsorption medium 88 desirable include a hydrophilic coatingor shell to provide biocompatibility, which is also desirablycharacterized by a high biocompatibility index. The coating material forthe particles 96 can be selected from the same group of materials as thecoating 80 for the particles 76 of the first adsorption medium 34.

In addition, the polymer particles 96 can also possess positivelycharged functional groups on the surface of the hydrophobic pores tofurther attract endotoxin through an ionic interaction. The amount ofthese positively charged groups desirably remains low, preferably below1 meq/ml. Thus, the overall hydrophobic nature of the core of thepolymeric particle is not compromised, so that hydrophobic interactionsstill remain the major mechanism of adsorption of LPS endotoxin. Thepositively charged functional groups covalently bonded to the surface ofthe pores of the polymeric particles 96 can be selected from the groupcomposed of amino-, methylamino-, ethylamino-, dimethylamino-,diethylamino-, ethanolamino-, diethanolamino-, polyethylenimino-groups,imidazole, histamine, or basic amino acids as lysine, arginine,histidine.

2. Removal of Other Materials

The adsorption medium 88 can also be composed to selectively adsorbother targeted proteins or toxins that can be released into the blood asa result of injury or trauma, e.g., myoglobin, which can be releasedduring a crush injury. The adsorption medium 88 can also be composed toselectively adsorb targeted chemical moieties that can be released intothe blood as a result of injury or trauma, e.g., potassium, which can bereleased with myoglobin during a crush injury.

The device 18 or 30 can also be used in combination with other devicesthat remove materials from the blood other than by selective adsorption,e.g., by ion exchange effects.

III. Systems and Methods for Removing Cytokines or Other Species ofPro-Inflammatory or Anti-Inflammatory Stimulators or Mediators fromPhysiologic Fluids

FIG. 14 shows an embodiment of a system 100 for removing cytokines orother species of pro-inflammatory or anti-inflammatory stimulators ormediators from a physiologic fluid. In this embodiment, the physiologicfluid comprises fresh peritioneal dialysis solution that has beenregenerated from spent peritoneal dialysis solution.

As shown in FIG. 14, the system 100 is configured for conducting a formof automated peritoneal dialysis. The system 100 includes a cycler 114,to automatically infuse, dwell, and drain peritoneal dialysis solutionto and from the patient's peritoneal cavity 120, typically at nightwhile the patient is asleep.

The system 100 includes a peritoneal dialysis solution flow set 112 thatestablishes communication between the system 100 and the peritonealcavity 120 of the patient. The cycler 114 interacts with the flow set112, to pump peritoneal dialysis solution into and out of the patient'speritoneal cavity 120 in prescribed infuse, dwell, and drain cycles.

The flow set 112 includes an in-line regeration module 122. The cycler114 circulates peritoneal dialysis solution, removed from the patient'speritoneal cavity 120, into the module 122 The cycler 114 alsocirculates a regeneration solution containing, e.g., electrolytes and/orbuffering materials, from a source 115 into the module 122.

The module 122 includes a component, e.g., a membrane, that transportswaste and uremic toxins from the spent peritoneal dialysis solution intothe regeneration solution, while also transporting electrolytes andbuffering materials from the regeneration solution 115 into theperitoneal dialysis solution. Typically, the regeneration fluid, ladenwith toxins and depleted of electrolytes and buffers, is sent to waste.

The module 122 thereby performs on-line regeneration of peritonealdialysis solution. Upon regeneration, the cycler 114 re-circulates theperitoneal dialysis solution back to the peritoneal cavity 120 of thepatient.

The spent peritoneal dialysis solution may carry cytokines or otherspecies of pro-inflammatory or anti-inflammatory stimulators ormediators generated while the solution dwelled within the peritonealcavity of the patient. Extracorporeal processing of the spent solutionby the cycler 114 can also trigger additional production of cytokines orother species of pro-inflammatory or anti-inflammatory stimulators ormediators.

The system 100 therefore includes a device 130 that removes cytokines orother species of pro-inflammatory or anti-inflammatory stimulators ormediators from the physiologic peritoneal dialysis solution prior to itsreturn to the patient's peritoneal cavity 120. The device 130'can becoupled to the system 100 either upstream or downstream of theregeneration module 122. In this arrangement, cytokines or other speciesof pro-inflammatory or anti-inflammatory stimulators or mediators areremoved from the peritoneal dialysis solution either before or afterregeneration, and prior to return to the regenerated solution to theperitoneal cavity 120 of the patient. This leads to overall reducedlevels of cytokines or other species of pro-inflammatory oranti-inflammatory stimulators or mediators in the peritoneal dialysispatient.

It should be appreciated that the device 122 can be used in otherperitoneal dialysis modalities where regeneration of peritoneal dialysissolution is performed.

Body fluids that are removed from and then recycled back to the bodyduring a given treatment modality can also carry cytokines or otherspecies of pro-inflammatory or anti-inflammatory stimulators ormediators, or cytokines or other species of pro-inflammatory oranti-inflammatory stimulators or mediators can be generated as a resultof such treatment modalities. Treatment systems and methods exist forremoving and recycling such fluids, e.g., lymphatic fluid, synovialfluid, spinal fluid, or cerebrospinal fluid. The devices, systems, andmethods that embody this aspect of the invention, as just discussed inthe context of peritoneal dialysis, can likewise be used in associationwith such treatment modalities, to remove cytokines or other species ofpro-inflammatory or anti-inflammatory stimulators or mediators from thebody fluids before, during, or after other forms of primary treatment.

FIG. 15 shows another embodiment of a system 200 for removing cytokinesor other species of pro-inflammatory or anti-inflammatory stimulators ormediators from a physiologic fluid. In this embodiment, the physiologicfluid comprises preservation solution 206 for a harvested organ 202awaiting transplantation.

As shown in FIG. 15, the system 200 includes a bath 204 holding theorgan 202. The preservation solution 206 is circulated from a source 208through the bath 204 and through the organ 202. FIG. 15 depicts aharvested kidney 202, but the organ can be any solid organ harvested fortransplant.

The organ 202 may generate cyctokines or other species ofpro-inflammatory or anti-inflammatory stimulators or mediators whileimmersed in the bath 204. The cytokines or other species ofpro-inflammatory or anti-inflammatory stimulators or mediators will, inturn enter the preservation solution 206 contacting and perfusing theorgan 202. Circulation of the preservation solution may also triggeradditional production of cytokines or other species of pro-inflammatoryor anti-inflammatory stimulators or mediators.

The system 200 therefore includes a device 230 that removes cytokines orother species of pro-inflammatory or anti-inflammatory stimulators ormediators from the preservation solution. The device 230 can be coupledto the system 200 either upstream or downstream of the bath 204. In thisarrangement, cytokines or other species of pro-inflammatory oranti-inflammatory stimulators or mediators are removed from thepreservation solution, so that the overall population of cytokines orother species of pro-inflammatory or anti-inflammatory stimulators ormediators to which the organ 202 is exposed prior to transplantation isminimized. This leads to overall reduced levels of cytokines or otherspecies of pro-inflammatory or anti-inflammatory stimulators ormediators in the patient who receives the organ transplant.

Either device 120 or 230 can be constructed in generally the samefashion already described with respect to devices 18 or 30.

EXAMPLE 1 Blood Purification Using an Adsorption Medium to RestoreImmunologic Stability

A study was conducted to demonstrate the ability of a biocompatibleadsorption medium to selectively adsorb cytokines (TNF, IL-6, and IL-10)from the blood. The medium comprised particles (as generally shown inFIG. 12) formed of a core of hydrophobic, crosslinked porousdivinylbenzene material coated with a thin, permeable biocompatiblehydrophilic polyvinylpyrrolidone material. The core material of theparticles possessed a mean pore size of about 16 nm. The particles werecontained within a housing (as generally shown in FIG. 3) and presenteda surface area to blood flow of about 650 sq.mg. The medium was obtainedfrom RenalTech International, New York, N.Y. (BetaSorb™ AdsorptionMedium).

The medium was tested in an experiment using in three animals subjectedto cecal ligation and puncture (CLP) 18 hrs earlier. The animalstolerated treatment with the medium without difficulty. The cytokineresponse was characterized over the four hours of treatment (see FIG.17).

The results demonstrate that the medium removed all three cytokines fromthe blood. As FIG. 17 shows, there was. a flattening out or evendownward trend in the concentrations of TNF, IL-6 and IL- 10 (in orderto keep the scales similar, the units for TNF in FIG. 17 are pg/ml, IL-6are ng/dl, and IL-10 are pg/cl). Previous experience with this model hasshown a progressive increases in IL-6 and IL-10 over a similar timeperiod and a more persistent TNF signal.

EXAMPLE 2 Biocompatibility Index of the Adsorption Medium

The adsorption medium employed in Example 1 was subjected to theprescribed battery of tests under the biocompatibility index testprotocol described above. The blood drawn from six individual healthydonors was subjected to the test protocol and the test results wereaveraged.

FIGS. 18A, 18B, and 18C show the average variations in blood cell countsfor red blood cells, white blood cells, and platelets, respectively,incrementally during passage of 25 ml of the blood through the treatmentdevice containing the medium. With respect to red blood cells, whiteblood cells, and platelets, the maximum difference between the base line(line S.K./A) and the medium (line S.K./B) was less than 15%.

FIG. 19 shows the average variations in PMN elastase concentrations(indicative of leukocyte activation) incrementally during passage of 25ml of the blood through the treatment device containing the medium. Themaximum difference between the based line (line S.K./A) and the medium(line S.K./B) was less than 15%.

FIG. 20 shows the average variations in LDH concentrations (indicativeof hemolysis) incrementally during passage of 25 ml of the blood throughthe treatment device containing the medium. The maximum differencebetween the based line (line S.K./A) and the medium (line S.K./B) wasless than 15%.

FIG. 21 shows the average variations in C3a-desArg concentrations(indicative of complement activation) incrementally during passage of 25ml of the blood through the treatment device containing the medium. Onedonor experienced a rapid increase in the C3a-desArg level from 86 up to822 μg/L due to clotting in the test system. The other five donors (whoexperienced no clotting in the test system) underwent more moderateincreases, with a mean increase of from 113 to 392 μg/L. The maximumdifference between the based line (line S.K./A) and the medium (lineS.K./B) was greater than 25%.

FIG. 22 shows the average variations in TAT concentrations (indicativeof coagulation) incrementally during passage of 25 ml of the bloodthrough the treatment device containing the medium. The maximumdifference between the based line (line S.K./A) and the medium (lineS.K./B) was less than 15%.

The following table lists the scoring the results for the indications asthe dimensionless quantities 1, 2, and 3.

Numeric Scores 1 2 3 (Signifying (Signifying (Signifying Most an a LeastDesired Acceptable Desired Blood Degree of Degree of Degree of IndicatorBiocompatibility) Biocompatibility) Biocompatibility) Loss/of 1 WhiteBlood Cells Loss of Red 1 Blood Cells Loss of 1 Platelets PMN Elastase 1Concentration LDH 1 Concentration C3a-desArg 3 Concentration TAT 1Concentration

The Biocompatibility Index for the Medium is 9, which indicates themedium can contact the blood with no significant loss of blood cells, nosignificant hemolysis, no significant activation of luekocytes ormonocytes, and, at most, only moderate complement activation, even withthe use of heparin as the sole anticoagulant. Because such materials arenot likely to induce the generation of cytokines, they are well suitedfor use to remove cytokines from the blood, blood products, orphysiologic fluids.

Various features of the invention are set forth in the following claims.

1. A method for treating an individual experiencing a condition on acontinuum from early sepsis to septic shock comprising the steps ofdrawing blood from the circulatory system of the individual for returnto the circulatory system, and removing cytokines from the blood bybringing the blood into contact with an adsorption medium sized toselectively adsorb cytokines from the blood, the adsorption mediumcomprising polymeric particles each comprising a hydrophobic core and abiocompatible hydrophilic coating, the material of the second unit beingcharacterized by a Biocompatibility Index of not greater than 14 derivedby a protocol consisting essentially of (i) selecting blood indicatorswhich quantify, physiologic changes based upon contact between theadsorption medium and blood, the blood indicators consisting essentiallyof (1) white blood cell count diminution as a result of contact with theadsorption medium ascertained by Coulter Counter; (2) red blood cellcount diminution as a result of contact with the adsorption mediumascertained by Coulter Counter; (3) platelet count diminution as aresult of contact with the adsorption medium ascertained by CoulterCounter; (4) leukocytes activation as a result of contact with theadsorption medium ascertained by measuring polymorphonuclear leukocyteelastase concentration (PMN Elastase Concentration); (5) complementactivation as a result of contact with the adsorption medium ascertainedby measuring anaphylatoxin C3 a-desArg concentrations; (6) occurrence ofhemolysis as a result of contact with the adsorption medium ascertainedby determining concentrations of Lactate dehydrogenase (LDH); andreduction of clot formation as a result of contact with the adsorptionmedium ascertained by measuring concentrations ofthrombin-antithrombin-complex (TAT); (ii) for each indicator,ascertaining a maximum difference between the indicator values over 25ml of flow of heperinized blood heperinized to a final concentration of1.0 IU heparin/ml blood passed through a biocompatible housing withoutthe adsorption medium, comprising a baseline value, and heparinizedblood passed through the housing containing the adsorption medium, andfor each indicator, expressing the maximum change as a percentagechange, relative to the baseline value; (iii) scoring the percentagechange for each indicator as a dimensionless numeric quantity 1, 2, or3, depending upon the magnitude of the percentage change, in accordancewith Table 1; and (iv) after scoring each indicator with a numericquantity of 1, 2, or 3, adding the numeric quantities scored for all theindicators to obtain a total, the total comprising the BiocompatibilityIndex.
 2. A method according to claim 1 wherein the BiocompatibilityIndex is not greater than
 7. 3. A method according to claim 1 whereinthe polymeric material comprises particles formed from a poroushydrophobic divinylbenzene copolymer having a surface modified toinclude surface exposed functional groups selected from the group ofpolymers of 2-hydroxyethyl methacrylate and N-vinylpyrrolidine.